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基于富含胺的碳点在均相体系中直接相互作用的酪氨酸酶活性荧光传感:检测机制与应用

Fluorescence sensing of tyrosinase activity based on amine rich carbon dots through direct interaction in a homogeneous system: detection mechanism and application.

作者信息

Zhang Jianzhong, Chen Yuyuan, Zheng Zongfu, Wang Zhenzhen, Zheng Yanjie, Lin Xinhua, Weng Shaohuang

机构信息

Department of Gastric Surgery, Fujian Medical University Union Hospital Fuzhou 350001 China.

Department of Pharmaceutical Analysis, The Higher Educational Key Laboratory for Nano Biomedical Technology of Fujian Province, School of Pharmacy, Fujian Medical University Fuzhou 350122 China

出版信息

RSC Adv. 2019 Jun 27;9(35):20029-20034. doi: 10.1039/c9ra03098k. eCollection 2019 Jun 25.

Abstract

As a vital, copper-containing oxidase, tyrosinase (TYR) is useful as a biomarker for the screening of skin diseases. In this paper, a convenient and sensitive homogeneous fluorescence detection platform for the assay of TYR activity without any modified steps is described. Inspired by the fact that carbon dots (CDs) with excellent properties can be obtained through some surface modification, amine rich carbon dots (N-CDs) using a nitrogen doping process were developed as the fluorescent probe for this assay. The effect and the response mechanism of the degree of nitrogen doping in relation to the response of different CDs to the sensing of TYR activity using dopamine (DA) as a substrate were investigated. The DA was oxidized to -dopaquinone with the catalyzation of TYR and quenched the fluorescence of the N-CDs by direct interaction. By using a set concentration of DA and other optimized reaction conditions, the fluorescence intensity of the N-CDs was directly applied to monitor the TYR activity. This assay for TYR activity showed a broad linear range from 0.05 to 6.0 U mL with a detection limit of 0.039 U mL. The satisfactory recovery of the sensor for TYR activity in diluted human serum illustrated a potential clinical application.

摘要

作为一种重要的含铜氧化酶,酪氨酸酶(TYR)可作为皮肤病筛查的生物标志物。本文描述了一种便捷、灵敏的均相荧光检测平台,用于检测TYR活性,无需任何修饰步骤。受通过一些表面修饰可获得具有优异性能的碳点(CDs)这一事实的启发,采用氮掺杂工艺开发了富胺碳点(N-CDs)作为该检测的荧光探针。研究了氮掺杂程度对不同CDs以多巴胺(DA)为底物检测TYR活性响应的影响及响应机制。在TYR的催化下,DA被氧化为多巴醌,并通过直接相互作用淬灭N-CDs的荧光。通过使用设定浓度的DA和其他优化的反应条件,N-CDs的荧光强度直接用于监测TYR活性。该TYR活性检测方法的线性范围为0.05至6.0 U/mL,检测限为0.039 U/mL。该传感器在稀释人血清中对TYR活性的回收率令人满意,表明其具有潜在的临床应用价值。

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